Abebe Genetu Bayih, PhD University of Gondar

Slides:

Advertisements

Similar presentations

MATERIALS AND METHODS Leaves and roots of Moringa stenopetala were collected around Arbaminch town (Southern Ethiopia) in October The plant parts.

Advertisements

Supplementary Figure S1.

Calcium dependent protein kinases and Malaria: Identification of chemical start point for drug discovery Symposium: Academic Drug Discovery: Challenges.

Focusing on design, experimental synthesis, model system, mechanization and results. BY ALEX PETER(A )

Malaria treatment. Dr abdulrahman al shaikh.. Introduction million patients died because of malaria every year. Most deaths due to Plasmodium Falciparum.

1 Discovering new drugs in Africa Defeating Malaria Together Kelly Chibale PhD FRSSAf University of Cape Town.

Identification of New Inhibitors of Plasmodium falciparum Enoyl- ACP Reductase Symposium on: Advances in Parasitology “Education and Research in Parasitology.

Isosteviol derivatives induced apoptosis in Human lung cancer via targeting MEK/MAPK pathway: An in vitro and in vivo study Ahmed M Malki 1,,PhD Stephen.

BLI Research Project By: Tiffany Hou Artemisinin.

Instituto de Medicina Molecular Lisboa, Portugal Miguel Prudêncio New drugs for treating and eradicating malaria.

Summary and Conclusion Hypothesis and Rationale

Novel Transcription Factor Inhibitor as Treatment for Epithelial Cell Cancers John Bushweller, Department of Molecular Physiology and Biological Physics,

Combination Therapy with Novel Nanoparticle, SNS01-T, and Lenalidomide Triggers Synergistic Cytotoxicity in Vitro and in Vivo in Multiple Myeloma and Mantle.

Farletuzumab in platinum sensitive ovarian cancer with low CA125

Fig. 1. APG increased the sensitivity of BEL-7402/ADM cells to ADM

Malaria Research in the Real World

A flavone-based inhibitor of in vitro human rhinovirus replication induces resistant mutations in capsid protein VP4. Céline Lacroixa, Hari Babu Bollikollac,

1. Abstract 3. Objective of the study 4. Methods 5. Results

Featured Inhibitors for October-BOC Sciences

…driving discovery An improved potent direct thrombin inhibitor shows efficacy with low bleeding risk Anirban Datta et al.

A novel approach to target fasting and post-prandial triglycerides

First insight into the effect of single oral dose therapy with artemisinin –naphthoquine phosphate combination in a mouse model of Schistosoma mansoni.

Efficacy Study Of Potential Anti-tubercular Molecules:

Enhancement of paclitaxel-mediated cytotoxicity in lung cancer cells by 17-allylamino geldanamycin: in vitro and in vivo analysis Dao M Nguyen, MD, Dominique.

S55746 synergizes with panobinostat in vitro and in vivo in DLBCL

In vitro and in vivo role of heat shock protein 90 in Amphotericin B resistance of Aspergillus terreus G. Blum, B. Kainzner, K. Grif, H. Dietrich, B.

Izabela Mlynarczuk-Bialy, Thorsten R

The impact of the IGF-1 system of cancer cells on radiation response – An in vitro study Senthiladipan Venkatachalam, Esther Mettler, Christian Fottner,

Volume 15, Issue 2, Pages (February 2007)

Marissa V. Powers, Paul A. Clarke, Paul Workman Cancer Cell

Volume 28, Issue 3, Pages (September 2015)

In vivo prophylactic and therapeutic efficacy of C12G6 in mice

LIMITATIONS OF CYTOTOXICITY assays

Fig. 6. Treatment with a DLK inhibitor is neuroprotective and reverses stress-induced gene expression changes. Treatment with a DLK inhibitor is neuroprotective.

Marissa V. Powers, Paul A. Clarke, Paul Workman Cancer Cell

The PI3K–Akt pathway is essential for the in vivo maintenance of murine Tregs. The PI3K–Akt pathway is essential for the in vivo maintenance of murine.

The Triterpenoid CDDO-Me Delays Murine Acute Graft-versus-Host Disease with the Preservation of Graft-versus-Tumor Effects after Allogeneic Bone Marrow.

Volume 22, Issue 6, Pages e4 (December 2017)

Survival of BALB/c mice after HK/Syd challenge.

Fig. 7 BRD0705 impairs colony formation in AML cell lines and patient cells and shows in vivo efficacy in multiple AML mouse models. BRD0705 impairs colony.

Fig. 4 DMF enhances VSVΔ51 therapeutic efficacy in syngeneic and xenograft tumor models. DMF enhances VSVΔ51 therapeutic efficacy in syngeneic and xenograft.

Volume 44, Issue 3, Pages (March 2016)

Volume 12, Issue 6, Pages (December 2012)

Oncolytic VSV Primes Differential Responses to Immuno-oncology Therapy

Volume 23, Issue 5, Pages (May 2016)

Volume 19, Issue 12, Pages (December 2012)

Volume 4, Issue 3, Pages (March 2015)

Fig. 8 SQLE inhibitor terbinafine suppresses NAFLD-HCC growth in vitro and in vivo. SQLE inhibitor terbinafine suppresses NAFLD-HCC growth in vitro and.

Volume 16, Issue 2, Pages (February 2009)

In vivo validation of effects of combination therapies on subpopulation composition. In vivo validation of effects of combination therapies on subpopulation.

Jae Won Chang, Daniel K. Nomura, Benjamin F. Cravatt

Volume 25, Issue 1, Pages (January 2017)

A and B, CO-1686–resistant NCI-H1975 cell clones, COR1-1 and COR10-1 display a reduced dependence on EGFR signaling for survival. A and B, CO-1686–resistant.

Volume 46, Issue 4, Pages (April 2017)

Effect of 14 hit compounds on SMN protein expression in type I SMA patient fibroblast cells (GM03813). Effect of 14 hit compounds on SMN protein expression.

PMPs survive STZ cytotoxicity and produce larger colonies with a higher yield of insulin+ progeny. PMPs survive STZ cytotoxicity and produce larger colonies.

Fig. 2. In vitro activity of DSM265 and its analogs on DHODH and P

Volume 17, Issue 8, Pages (August 2010)

GR cells are dependent upon sustained CDC25C signaling as pharmacologic or genetic inhibition of CDC25C induce synthetic lethality. GR cells are dependent.

PTENP1 sensitizes renal cancer cells to chemotherapy.

Treatment effects of fenretinide (4-HPR, 5 μmol/L), 2-ME (2

Pretreatment of donor T cells with a c-Rel antagonist does not impair GVT activity. Pretreatment of donor T cells with a c-Rel antagonist does not impair.

A to C, MetMAb shows strong antitumor activity in the KP4 orthotopic model of pancreatic cancer by ultrasound. A to C, MetMAb shows strong antitumor activity.

In vivo prophylactic and therapeutic efficacy of C12G6 in mice

Efficacy of KM100 and/or MTX in BALB/c mice bearing subcutaneous TUBO tumors. Efficacy of KM100 and/or MTX in BALB/c mice bearing subcutaneous TUBO tumors.

Flavopiridol inhibits rhabdoid cell survival and induces cell cycle arrest and apoptosis. Flavopiridol inhibits rhabdoid cell survival and induces cell.

IL-1β expression is increased in MM cell lines by the co-culture with platelets in vitro. IL-1β expression is increased in MM cell lines by the co-culture.

Primary screening results of the MMV Malaria Box library.

BH3-targeted inhibitors drive specific resistance in human cell lines, which can be overcome with alternating or combining inhibitors. BH3-targeted inhibitors.

Knockdown of ROR1 increases the invasive potential of melanoma cells in vitro and in vivo. Knockdown of ROR1 increases the invasive potential of melanoma.

Presentation transcript:

Abebe Genetu Bayih, PhD University of Gondar In vitro and in vivo antimalarial activity of novel harmine-analog heat shock protein 90 inhibitors: a possible partner for artemisinin. Abebe Genetu Bayih, PhD University of Gondar

Introduction Malaria In 2015: 3.2 billion people at risk Source: WHO Malaria In 2015: 3.2 billion people at risk 214 million cases 438,000 deaths 90% in sub-Saharan Africa Children most affected (WHO 2015 report) Significant reduction in mortality and morbidity Emergence of ART resistance – a challenge! Catastrophe if reaches Africa! NEW DRUGS NEEDED!

Heat shock proteins (Hsp): Introduction… Heat shock proteins (Hsp): Molecular chaperone Function – protein folding, intracellular trafficking, gene expression, cell cycle, and cell differentiation. Conserved – from bacterial to mammals Constitutive and induced Temperature, nutrition, heavy metal exposure antimalarial and anti-cancer drug target Hsp90 Structure (Bracher and Hartl, 2006)

Introduction… PfHsp90 inhibitors: Hypothesis: Natural compounds (Geldanamycin and derivatives) (Kumar et al, 2003; Banumathy et al, 2003) Harmine (Shahinas et al, 2010, 2012) Beta-carboline alkaloid Preferentially binds to the ATP-binding domain of PfHsp90 Inhibits P. falciparum in vitro Antimalarial effect in vivo in mice Hypothesis: Structural modification of harmine improves efficacy. Harmine

METHOD 45 novel harmine analogs screened for PfHsp90 binding Two (17A and 21A) bind to PfHsp90 Cytotoxicity assay (HepG2 and HeLa) In vitro antimalarial assay (P. falciparum) Fixed dose in vivo antimalarial assay (P. berghei ANKA in BALB/c mice) 21A - in vivo dose-ranging expt. 21A and DHA in vivo combination expt.

RESULTS AND DISCUSSION 1. Competitive binding assay bis-ANS assay: Mean EC50 ± SEM (µM) : 17A = 12.2 ± 2.3 ; 21A = 23.1 ± 8.8; Radicicol = 5.46 ± 2.42 2. In vitro antimalarial activity 72h in vitro susceptibility assay HRP-II ELISA Mean IC50 ± SEM (µM) : PF-W2: 17A = 4.2 ± 1.3 21A = 5.7 ± 1.7 CQ = 0.15 ± 0.04 PF-3D7: 21A = 13.5 ± 0.8 PF-MRA 1236: 21A = 9.2 ± 0.4 PF-MRA 1240: 21A = 9.6 ± 2.0

3. In vivo antimalarial activity Fixed-dose harmine-derivatives against P. berghei in BALB/c mice (100mg/kg) I. Parasitemia: At day-7 post-infection: Percent inhibition: 17A = 51.5% 21A = 56.1% 17A and 21A significantly lower parasitemia than vehicle ctrl. (p<0.05).

II. Survival Drug treated mice showed better survival than the vehicle group (DMSO). The Median Survival time: 17A = 11, 21A = 14d, DMSO = 8.5d. 21A resulted higher survival than 17A Significant difference b/n 21A and DMSO (Log-rank (Mantel-Cox) test, p=0.0177)

B. Dose-ranging experiment 21A Parasitemia: Percent parasitemia inhibition 100 mg/kg 75 50 25 D6 48.1 37.5 27.2 18.4 D7 45.9 33.4 18.9 At D6, treatment with 100mg/kg, 75mg/kg and 50mg/kg significantly reduced parasitemia. The Median Survival time: 100mg/kg = 11.5d, 75mg/kg = 10d, 50mg/kg = 10d, 25mg/kg = 8d, DMSO = 8d.

C. 21A-DHA combination experiment 21A (100mg/kg) showed additive effect with DHA (10mg/kg) Parasite clearance: One day after two doses of drug: 21A = 0/5 (0%) DHA = 2/5 (40%) 21A plus DHA = 5/5 (100%) One day after three doses of drug: DHA = 3/5 (60%) A combination of 21A and DHA improved survival rate.

4. Cytotoxicity CCK-8 (Sigma): Based on water soluble WST-8 Live/dead cell Mean IC50 ± SEM (mM) : HepG2: 17A = 0.31 ± 0.023 21A = 0.093 ± 0.002 Geldanamycin = 0.00038 ± 8.98E-05 Chloroquine = 0.24 ± 0.024 HeLa: 17A = 0.738 ± 0.14 21A = 0.436 ± 0.089 17A and 21A NOT toxic to HepG2 and HeLa cells.

SUMMARY Out of 45 harmine analogs tested, 17A and 21A: Bind to ATP-binding domain of PfHsp90 effectively. Inhibit P. falciparum in vitro in micromolar concentration Significantly reduce parasitemia in vivo Prolong survival of infected mice (not seen in parent molecule, Harmine) 21A showed a dose-dependent antimalarial effect. 21A showed additive effect with DHA Not toxic in HepG2 and HeLa cells in vitro 21A could be a potential standalone or a partner antimalarial drug.

Acknowledgement: Asongna Folefoc, PhD, University of Calgary, Canada Abu Naser Mohon, MSc, University of Calgary, Canada Scott Eagon, PhD, San Francisco State University, USA Marc Anderson, PhD, San Francisco State University, USA Dylan R. Pillai, MD, PhD, University of Calgary, Canada Funding: Grandchallenges Canada to DRP

Towards a malaria-free happy world!